| 刘 超,刘召波,刘海东,吴 敏,张爱英,李 宁.AFP-L3蛋白芯片的建立与应用[J].现代生物医学进展英文版,2018,(15):2977-2981. |
| AFP-L3蛋白芯片的建立与应用 |
| Development and Application of Protein Microarray for Detecting Alpha-fetoprotein-L3 |
| Received:January 03, 2018 Revised:January 25, 2018 |
| DOI：10.13241/j.cnki.pmb.2018.15.040 |
| 中文关键词: AFP-L3 蛋白芯片 ELISA 凝集素微量离心柱法 肝细胞癌 诊断 |
| 英文关键词: AFP-L3 Protein microarray ELISA Lens culinaris agglutinin-coupled spin column Hepatocellular carcinoma Diagnosis |
| 基金项目:国家"十二五"科技成果转化立项项目(2015ZX10004801)子课题；国家艾滋病和病毒性肝炎等重大疾病传染病防治科技重大专项(2012ZX10002017-007)；肝癌抗复发转移治疗临床新体系的研究和应用推广；北京市属医学科研院所公益发展改革试点项目(京医研2016-2)；肝癌特异性早期预测和分期诊断分子标志物系统筛选及机制研究平台；北京市科委课题(D121100003912002)；建立乙型肝炎肝硬化无创诊断标准及模型的研究；北京市卫生系统高层次卫生技术人才培养计划(2013-3-074)；北京市教育委员会项目(KM201510025020)；北京市教委重大传染病防治协同创新中心；北京市医院管理局重点医学专业发展计划(ZYLX201311)；2017北京市肝病研究所自主课题项目，乙肝相关肝癌血清蛋白标志物蛋白芯片的研发与临床验证 |
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| 中文摘要: |
| 摘要 目的：建立一种可同时检测血清中甲胎蛋白(AFP)及甲胎蛋白异质体(AFP-L3)的蛋白芯片方法，为AFP-L3检测提供经济、便捷、省时、有效的新途径。方法：将鼠源AFP单克隆抗体和小扁豆凝集素点样固定在醛基玻片上，制备出AFP抗体和小扁豆凝集素蛋白芯片。利用抗原抗体特异性结合以及盐藻糖与小扁豆凝集素特异性结合的原理，用蛋白芯片方法检测血清样本中的AFP和AFP-L3。结果：肝癌血清39份，其中37份检测到AFP；26份肝癌血清中同时检测到AFP和AFP-L3，2份肝癌血清均未检测到AFP和AFP-L3。肝细胞癌组血清样本中AFP和AFP-L3水平明显高于健康对照组(P<0.001)，健康对照组与空白对照组统计学上无差异(P>0.05)。结论：本研究成功地建立AFP和AFP-L3同时检测的蛋白芯片方法，与ELISA法和凝集素微量离心柱法相比，是一种切实可行，经济、便捷、省时、有效的方法。 |
| 英文摘要: |
| ABSTRACT Objective: To develop a protein microarray method for detecting AFP and AFP-L3 in serum samples at the same time and provide an economical, feasible, time-saving and efficiency method for screening of primary hepatic carcinoma. Methods: Spotting mouse-derived AFP monoclonal antibody and Lens culinaris agglutinin on an aldehyde glass slide to develop a protein microarray method for detecting AFP and AFP-L3 of HCC patients and healthy controls. The protein microarray was based on the antibody-antigen reaction principle and Lens culinaris agglutinin-Fucose reaction principle. Results: AFP was detected in 37 cases of 39 HCC samples. AFP and AFP-L3 were both detected in 26 cases of 37 HCC samples. AFP and AFP-L3 were both undetected in 2 cases of 39 HCC sam- ples. The AFP and AFP-L3 levels in serum of HCC patients were significantly higher than those of healthy controls(P<0.001). There was no statistically significant difference between healthy controls and blank controls (P>0.05). Conclusion: A protein microarray which can assay AFP and AFP-L3 at same time was successfully established. Compared with Lens culinaris agglutinin-coupled spin column(ACSC) technology and ELISA, it was a feasible, economical, convenient use, time-saving and efficiency method. |
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